黄芪升麻柴胡治疗实验性自身免疫性重症肌无力大鼠的量效关系及免疫学机制研究  

Research on dose effect relationship and immunological mechanism of astragalus,rhizoma and radix bupleuri in treatment of experimental autoimmune myasthenia gravis rats

作　　者：何骁隽 吴颢昕[1] 吴周烨 许梦贤 许骏尧 陈刚[1] 陶伟伟[1] HE Xiao-Jun;WU Hao-Xin;WU Zhou-Ye;XU Meng-Xian;XU Jun-Yao;CHEN Gang;TAO Wei-Wei(Basic Medical College of Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学基础医学院,南京210023

出　　处：《中国免疫学杂志》2018年第9期1336-1343,共8页Chinese Journal of Immunology

基　　金：江苏高校优势学科建设工程资助项目(No.161014);孟景春全国名老中医药专家传承工作室建设项目(No.013007001009)

摘　　要：目的:探讨益气升提法治疗实验性自身免疫性重症肌无力(EAMG)大鼠中黄芪的最佳剂量,以及免疫机制。方法:采用Rα97-116免疫Lewis大鼠构建EAMG,随机分为佐剂对照组、模型对照组、益气升提黄芪低剂量组、益气升提黄芪中剂量组、益气升提黄芪高剂量组。观察大鼠抓力、体质量、低频重复电刺激(RNS)衰减率,检测血清AChR-Ab、IFN-γ、TGF-β、IL-6、IL-17含量及外周血淋巴细胞CD4^+ CD25^+ Fox P3^+ Treg比例,将EAMG大鼠临床表现与实验室检测指标综合汇总,进行综合评分,评价各组治疗效果。结果:治疗后,低中高三剂量益气升提组大鼠EAMG综合评分分别为82,58,88分(满分100分),各组大鼠体质量回升,抓力回升,RNS衰减率显著降低(P<0. 05,P<0. 01,P<0. 001),血清AChR-Ab、IFN-γ、IL-6、IL-17含量显著下降,TGF-β含量显著升高(P<0. 05,P<0. 01,P<0. 001),外周血淋巴细胞CD4^+ CD25^+ Fox P3^+ Treg比例显著升高(P <0. 05)。结论:益气升提法对EAMG大鼠具有明确治疗效果,黄芪高剂量组疗效最优,不同剂量黄芪配伍升麻柴胡对MG细胞因子网络的影响各不相同。益气升提法治疗EAMG的机制可能是通过升高TGF-β含量,降低IFN-γ、IL-6、IL-17含量,提高CD4^+ CD25^+ Fox P3^+ Treg比例,降低血清AchR-Ab含量,从而减少神经肌肉接头处AchR损害。Objective:To investigate the dose effect relationship and the immune mechanism of astragalus in invigorating qi for ascending method in experimental autoimmune myasthenia gravis(EAMG)rats.Methods:EAMG models were constructed on Lewis rats with Rα97-116,randomly divided into 5 groups:CFA group,EAMG model group,invigorating qi for ascending astragalus low dose group,invigorating qi for ascending astragalus middle dose group and invigorating qi for ascending astragalus high dose group.The holding power,the weight and the decrement percentage of RNS were evaluated.ELISA was adopted to determine the titers of AchR-Ab,IFN-γ,TGF-α,IL-6 and IL-17 in serum.FCM was adopted to determine the proportion of CD4+CD25+FoxP3+Treg in peripheral blood lymphocytes.The EAMG comprehensive score was assessed according to the clinical symptoms and experimental analysis,which was used to evaluate the curative effect of each group.Results:After medication,the EAMG comprehensive score of each Chinese medicine group was--low dose group 82;middle dose group 58;high dose group 88(full score 100),the percentage of decrement of RNS of rats in three Chinese medicine groups and positive control group went down significantly,and there was an improvement of body weight and holding power(P<0.05,P<0.01,P<0.001);the level of AChR-Ab,IFN-γ,IL-6 and IL-17 descended significantly(P<0.01,P<0.05),with TGF-αlevel going up significantly(P<0.05,P<0.01,P<0.001);the proportion of CD4+CD25+FoxP3+Treg in peripheral blood lymphocytes increased significantly(P<0.05).Conclusion:Invigorating qi for ascending method has a significant therapeutic effect on MG.High dose of astragalus does the best.Different doses of astragalus combined with rhizoma and radix bupleuri has different effects on MG cytokine network.The immune mechanism of treatment may be inhibiting the synthesis of AChR-Ab or promoting its degradation by regulating IFN-γ,IL-6,IL-17 and TGF-αlevel and proportion of CD4+CD25+FoxP3+Treg,so that the damage of AchR of NMJ will be reduced.

关 键 词：益气升提法 实验性自身免疫性重症肌无力 重症肌无力综合评分 黄芪 量效关系 淋巴细胞亚群 免疫机制 

分 类 号：R285[医药卫生—中药学] R392[医药卫生—中医学]